Mammalian cell line models and related methods
a cell line model and mammalian technology, applied in the field of mammalian cell line models, can solve the problems of wasting time, unable to meet the needs of patients, so as to improve the characteristics of the cell line, improve the growth rate, and improve the effect of product production
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example i
Metabolic Model for NS0 Cell Line and Optimization of Media Formula
[0188]This example describes metabolic network reconstruction of the NS0 cell line.
[0189]A metabolic model for NS0 cell line was reconstructed and used to optimize media formulation in a GS-NS0 murine myeloma cell line, which expresses a human anti-rabies IgG monoclonal antibody. Although the model-based media optimization approach has been exemplified using an NS0 metabolic model and NS0 cell culture experiments, this approach is equally applicable to other cell lines, including but not limited to human- or animal-derived cell lines, for example, Chinese Hamster Ovary (CHO), BHK, NS0, SP2 / 0, 3T3, Hybridoma, C127, HEK293, PER.C6, HepG2, HeLa, MRC5, WI38, MDCK, Vero, COS, or modified cell lines such as GS-NS0 or CHO DHFR−. Such cell lines can be used, for example, for production of biologics, including but not limited to growth factors, monoclonal antibodies, hormones, cytokines, fusion proteins, recombinant enzymes, ...
example ii
Precursor Metabolite, Energy, and Biomass Synthesis in the Reconstructed Metabolic Model of NS0 Cell Line
[0200]This example describes characteristics of the reconstructed metabolic model of the NS0 cell line.
[0201]To assess the network's ability to synthesize biomass components, precursor metabolite formation and energy (ATP) production were simulated using glucose as the sole carbon source. The reconstructed network was able to correctly generate all the precursor metabolites at levels equal or below the maximum theoretical values from glucose, as it was expected and similar to previously reconstructed models for microbial cells such as E. coli and S. cerevisiae (Forster et al., Genome Res 13:244-253 (2003); Varma et al., Appl. Environ. Microbiol. 59:2465-2473 (1993)). In addition, using a P / O ratio of 2.5 (Sheikh et al., Biotechnol Prog. 21:112-121 (2005); Vo et al., J. Biol. Chem. 279, 39532-39540 (2004)), the metabolic model simulated ATP formation at a maximum yield of 32 mol A...
example iii
Metabolic Growth Simulation
[0206]This example describes simulation of metabolic growth for the NS0 cell line.
[0207]To simulate growth in the reconstructed model and to validate the accuracy of the network content, metabolic network parameters including the growth and non-growth associated energy requirements were determined using experimental data, and the model predictions were validated by comparing the simulation results to experimental measurements.
[0208]Metabolite Uptake and (By)product Secretion Rate Constraints in the NS0 Metabolic Model. To simulate cell physiology in a bioreactor, metabolite uptake and secretion rates and protein and biomass formation rates were calculated from cell culture data, as described above. The maximum uptake of input nutrients, or the maximum limit on nutrient uptake constraints in the model, was set to the rates calculated from the experimental data and the minimum values were constrained to zero, so the uptake of nutrients could vary between zer...
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